In vivo Imaging of Fully Active Brain Tissue in Awake Zebrafish Larvae and Juveniles by Skull and Skin Removal

作者： Paul Schramm1, Florian Hetsch2, Jochen C. Meier3, Reinhard W. Köster1 1Cellular and Molecular Neurobiology, Zoological Institute,Technische Universität Braunschweig, 2Institute of Pathophysiology,University Medical Center of the Johannes Gutenberg University, 3Cell Physiology, Zoological institute,Technische Universität Braunschweig

简介：

Overview

This study presents a microinvasive method for imaging the zebrafish embryonic brain in vivo, adaptable to larval and juvenile stages. By employing this technique, researchers can access detailed cellular and subcellular structures of mature neurons, facilitating the investigation of brain function and neuropharmacological responses.

Key Study Components

Area of Science

• Neuroscience
• Developmental Biology
• Optogenetics

Background

• Imaging the zebrafish brain in later larval stages offers previously unattainable views of neuronal architecture.
• Pigment cells that interfere with imaging can be removed using this method.
• The study focuses on examining synaptic plasticity, neuron degeneration, and regeneration processes.
• This technique is vital for investigating complex behaviors such as social interaction and decision-making in zebrafish.

Purpose of Study

• To present a method that allows high-resolution imaging of the zebrafish brain during larval stages.
• To facilitate studies on cellular processes affecting brain development and function.
• To contribute to a better understanding of social behavior and decision-making mechanisms.

Methods Used

• This method utilizes sharp, thin glass needles for microsurgery on zebrafish larvae.
• Zebrafish are chosen as the biological model due to their transparency and accessibility for imaging.
• The procedure involves anesthetizing the larvae and using low melting agarose for embedding.
• Imaging is conducted with an inverted microscope following an open skull surgery technique.
• Neuronal morphology and synaptic structures can be examined in larvae older than seven days post-fertilization.

Main Results

• The method enhances the visibility of brain structures by eliminating barriers such as skin and melanin.
• It allows for detailed imaging of neuronal morphology and intracellular processes.
• Blood vessels within the brain were visualized at depths of nearly 250 microns, showcasing the technique's effectiveness.
• Key findings suggest this approach can lead to new insights into the mechanisms underlying complex neural behaviors.

Conclusions

• This study demonstrates a technique that significantly improves the accessibility and imaging quality of zebrafish brain tissues.
• It allows for detailed examinations of neuromorphological changes and cellular processes during critical developmental stages.
• These advancements hold substantial implications for understanding the neural basis of behavior and potential interventions in neuronal processes.

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